Patterned, Responsive Cellular Therapies Using Novel Mammalian Cellular Regulator Systems

We propose to design, build and test genetic circuits and DNA constructs that will be expressed in human cells and that will ultimately have applications for the health of warfighters. The circuits will have physiological inputs representing, for example, (1) infection/sepsis, (2) altitude sickness or blood loss, and (3) radiation exposure; which will be mediated through signal transduction pathways initiated by inflammatory cytokines, hypoxia, and DNA damage. The circuit outputs will be secreted proteins that are adaptive for each of these physiological states, such as (a) antibodies that inhibit damaging cytokines and (b) pro-survival proteins that prevent cell death in response to trauma (particularly neuronal death). The DNA constructs will contain multiple circuits that will be activated independently, using distinct promoters, enhancers, and insulating elements to insure functional separation. The DNA constructs will be designed for administration to warfighters either by gene therapy or withdrawal of cells, engineering the cells in vitro, and return of engineered cells into the warfighter. The constructs will also contain circuits that allow for killing of engineered cells after their utility has expired. The constructs will not integrate into the genome and either not replicate at all or replicate only during phases of need.